JP2016521574A - Process for fermenting a CO-containing gaseous substrate in a low phosphate medium effective in reducing water use - Google Patents
Process for fermenting a CO-containing gaseous substrate in a low phosphate medium effective in reducing water use Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 56
- 230000008569 process Effects 0.000 title claims abstract description 52
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 51
- 239000000758 substrate Substances 0.000 title claims abstract description 46
- 229910019142 PO4 Inorganic materials 0.000 title claims abstract description 25
- 239000010452 phosphate Substances 0.000 title claims abstract description 25
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 title claims abstract description 24
- 238000000855 fermentation Methods 0.000 claims abstract description 99
- 230000004151 fermentation Effects 0.000 claims abstract description 99
- 229910052723 transition metal Inorganic materials 0.000 claims abstract description 11
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 229910052751 metal Inorganic materials 0.000 claims abstract description 8
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 238000001556 precipitation Methods 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 122
- 241000193403 Clostridium Species 0.000 claims description 26
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 24
- 239000000203 mixture Substances 0.000 claims description 15
- 241001656809 Clostridium autoethanogenum Species 0.000 claims description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
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- 229910052711 selenium Inorganic materials 0.000 claims description 7
- 229910052725 zinc Inorganic materials 0.000 claims description 7
- 241000193401 Clostridium acetobutylicum Species 0.000 claims description 6
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 6
- 241000204649 Thermoanaerobacter kivui Species 0.000 claims description 6
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
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- 241001534860 Alkalibaculum bacchi Species 0.000 claims description 4
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- 101000941690 Homo sapiens Cytochrome P450 1A1 Proteins 0.000 claims description 4
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- 230000002378 acidificating effect Effects 0.000 claims description 4
- 241001223493 Acetoanaerobium noterae Species 0.000 claims description 3
- 241001468163 Acetobacterium woodii Species 0.000 claims description 3
- 241001058118 Caldanaerobacter Species 0.000 claims description 3
- 241001157085 Caldanaerobacter subterraneus Species 0.000 claims description 3
- 241000620137 Carboxydothermus hydrogenoformans Species 0.000 claims description 3
- 241001656810 Clostridium aceticum Species 0.000 claims description 3
- 241001171821 Clostridium coskatii Species 0.000 claims description 3
- 241000328950 Clostridium drakei Species 0.000 claims description 3
- 241001468167 Clostridium magnum Species 0.000 claims description 3
- 241000193469 Clostridium pasteurianum Species 0.000 claims description 3
- 241000186587 Clostridium scatologenes Species 0.000 claims description 3
- 102100030787 ERI1 exoribonuclease 2 Human genes 0.000 claims description 3
- 241000186398 Eubacterium limosum Species 0.000 claims description 3
- 241001494297 Geobacter sulfurreducens Species 0.000 claims description 3
- 101000938751 Homo sapiens ERI1 exoribonuclease 2 Proteins 0.000 claims description 3
- 241000193459 Moorella thermoacetica Species 0.000 claims description 3
- 241001509483 Oxobacter pfennigii Species 0.000 claims description 3
- 241000191758 [Clostridium] ultunense Species 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 241000205275 Methanosarcina barkeri Species 0.000 claims 4
- 240000008067 Cucumis sativus Species 0.000 claims 2
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 2
- 244000061457 Solanum nigrum Species 0.000 claims 2
- 229910052698 phosphorus Inorganic materials 0.000 claims 2
- 239000011574 phosphorus Substances 0.000 claims 2
- 229910052700 potassium Inorganic materials 0.000 claims 2
- 239000011591 potassium Substances 0.000 claims 2
- 241001202853 Blautia Species 0.000 claims 1
- 229910052755 nonmetal Inorganic materials 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 114
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 87
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- 239000002609 medium Substances 0.000 description 55
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- 244000005700 microbiome Species 0.000 description 11
- 239000011550 stock solution Substances 0.000 description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000002309 gasification Methods 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 6
- 229910052739 hydrogen Inorganic materials 0.000 description 5
- 229910021654 trace metal Inorganic materials 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
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- 241000186566 Clostridium ljungdahlii Species 0.000 description 2
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
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- 229910021529 ammonia Inorganic materials 0.000 description 2
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- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000620141 Carboxydothermus Species 0.000 description 1
- 241001611022 Clostridium carboxidivorans Species 0.000 description 1
- 241001611023 Clostridium ragsdalei Species 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 241000592830 Desulfotomaculum kuznetsovii Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241000205276 Methanosarcina Species 0.000 description 1
- 241000178985 Moorella Species 0.000 description 1
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- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- 210000001822 immobilized cell Anatomy 0.000 description 1
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- 239000002244 precipitate Substances 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
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- 150000003722 vitamin derivatives Chemical class 0.000 description 1
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- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/065—Ethanol, i.e. non-beverage with microorganisms other than yeasts
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
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Abstract
CO含有ガス状基質を低ホスファート培地中で発酵させるためのプロセスを提供する。本プロセスは、少なくとも1種の遷移金属元素を含む液体培地を、少なくとも1種の他の遷移金属元素及び1種の非金属元素を含む液体培地とブレンドして発酵培地を与える工程を含む。本プロセスは、1種以上の遷移金属元素と1種以上の非金属元素との沈殿を防止するのに有効である。本プロセスに用いる発酵培地は、顕著に少ない量の水及び低減したレベルのホスファートを必要とするように調製される。【選択図】図1A process for fermenting a CO-containing gaseous substrate in a low phosphate medium is provided. The process includes blending a liquid medium containing at least one transition metal element with a liquid medium containing at least one other transition metal element and one non-metal element to provide a fermentation medium. This process is effective in preventing precipitation of one or more transition metal elements and one or more non-metallic elements. The fermentation medium used in this process is prepared to require a significantly lower amount of water and reduced levels of phosphate. [Selection] Figure 1
Description
この出願は、2013年6月10に出願され、その内容全体を参照によってここに援用する米国仮出願第61/833,240号の利益を主張する。
低ホスファート培地中でCO含有ガス状基質を発酵させるためのプロセスを提供する。さらに詳細には、本プロセスは、より少ない量の水を必要とするように調製された培地中でCO含有ガス状基質を発酵させる工程を含む。
This application claims the benefit of US Provisional Application No. 61 / 833,240, filed June 10, 2013, the entire contents of which are incorporated herein by reference.
A process is provided for fermenting a CO-containing gaseous substrate in a low phosphate medium. More particularly, the process includes fermenting a CO-containing gaseous substrate in a medium that is prepared to require a smaller amount of water.
背景
酢酸生成微生物は、ガス状基質の発酵を介して一酸化炭素(CO)からエタノールを生成することができる。クロストリジウム属由来の嫌気性微生物を用いる発酵は、エタノール及び他の有用な生成物を生成する。例えば、米国特許第5,173,429号は、合成ガスからエタノール及びアセタートを生成する嫌気性微生物であるクロストリジウム・リュングダリイ(Clostridium ljungdahlii)ATCC No. 49587について記載している。米国特許第5,807,722号は、クロストリジウム・リュングダリイATCC No. 55380を用いて廃ガスを有機酸とアルコールに変換するためのプロセス及び装置について記載している。米国特許第6,136,577号は、クロストリジウム・リュングダリイATCC No. 55988及び55989を用いて廃ガスをエタノールに変換するためのプロセス及び装置について記載している。
発酵プロセスは、大量の水と栄養素を必要とすることが多い。アルコールの生産性を維持しながら水の利用を低減し、特定成分を排除し、かつ他の成分の必要濃度レベルを低減すると、特に商業規模の発酵で顕著なコスト削減をもたらすことができる。
Background Acetic acid producing microorganisms can produce ethanol from carbon monoxide (CO) via fermentation of a gaseous substrate. Fermentation with anaerobic microorganisms from the genus Clostridium produces ethanol and other useful products. For example, US Pat. No. 5,173,429 describes Clostridium ljungdahlii ATCC No. 49587, an anaerobic microorganism that produces ethanol and acetate from synthesis gas. U.S. Pat. No. 5,807,722 describes a process and apparatus for converting waste gas to organic acids and alcohols using Clostridium lungdalii ATCC No. 55380. U.S. Pat. No. 6,136,577 describes a process and apparatus for converting waste gas to ethanol using Clostridium lünddalii ATCC No. 55988 and 55989.
Fermentation processes often require large amounts of water and nutrients. Reducing water utilization while maintaining alcohol productivity, eliminating certain components, and reducing the required concentration levels of other components can result in significant cost savings, especially in commercial scale fermentations.
概要
より少ない量の水を用いてCO含有ガス状基質を発酵させるプロセスを提供する。本プロセスに用いる発酵培地は、顕著に少ない量の水と低減したレベルのホスファートを必要とするように調製される。
SUMMARY A process for fermenting a CO-containing gaseous substrate using a smaller amount of water is provided. The fermentation medium used in this process is prepared to require a significantly lower amount of water and reduced levels of phosphate.
発酵プロセスは、少なくとも1種の遷移金属元素を含む液体培地を、少なくとも1種の他の遷移金属元素及び1種の非金属元素を含む液体培地とブレンドして発酵培地を与える工程を含む。本プロセスは、CO含有基質を発酵培地と接触させる工程及びCO含有基質を発酵させて酸性pHを与える工程を含む。本プロセスは1種以上の遷移金属元素と1種以上の非金属元素との沈殿を防止するのに有効であり、かつ生成されるエタノール1米国ガロン(3.8リットル)当たり発酵培地に与えられる約2米国ガロン(7.6リットル)以下の水を利用するのに有効である。
CO含有基質の発酵プロセスは、CO含有基質を発酵槽に与える工程及びCO含有基質を発酵培地と接触させる工程を含む。本プロセスは、Zn、Co及びNiから成る群より選択される1種以上の元素を含む第1溶液を、W及びSeから成る群より選択される1種以上の元素を含む第2溶液と、30mS/cm以下の伝導率及び約3mM以下のホスファートを有する発酵培地を与えるのに有効な量でブレンドする工程を含む。CO含有基質を発酵させる工程は、10gの全アルコール/(L・日)以上のSTYを与えるのに有効であり、かつ生成されるエタノール1米国ガロン(3.8リットル)当たり発酵培地に与えられる約2米国ガロン(7.6リットル)以下の水を利用するのに有効である。
別の態様では、発酵培地の調製における水の利用を低減するためのプロセスは、Zn、Co、Niの1種以上から成る群より選択される元素を含む溶液を、W、Seの1種以上から成る群より選択される元素を含む溶液と、約30mS/cm以下の伝導率を有する発酵培地を与えるのに有効な量でブレンドする工程を含む。この発酵培地は、約3mMより多くのホスファートを有する発酵培地より約10%〜約40%少ない水を必要とする。
本プロセスのいくつかの態様の上記及び他の態様、特徴及び利点は下記図面からさらに明らかになる。
The fermentation process includes blending a liquid medium containing at least one transition metal element with a liquid medium containing at least one other transition metal element and one non-metal element to provide a fermentation medium. The process includes contacting a CO-containing substrate with a fermentation medium and fermenting the CO-containing substrate to provide an acidic pH. The process is effective in preventing precipitation of one or more transition metal elements and one or more non-metallic elements, and about 2 fed to the fermentation medium per 1 US gallon (3.8 liters) of ethanol produced. It is effective to use water less than US gallon (7.6 liters).
The fermentation process for the CO-containing substrate includes the steps of providing the CO-containing substrate to a fermentor and contacting the CO-containing substrate with a fermentation medium. The process includes a first solution containing one or more elements selected from the group consisting of Zn, Co, and Ni, a second solution containing one or more elements selected from the group consisting of W and Se, Blending in an amount effective to provide a fermentation medium having a conductivity of 30 mS / cm or less and a phosphate of about 3 mM or less. The process of fermenting the CO-containing substrate is effective to give 10 g total alcohol / (L · day) or more STY, and about 2 fed to the fermentation medium per 1 US gallon (3.8 liters) of ethanol produced. It is effective to use water less than US gallon (7.6 liters).
In another aspect, the process for reducing the use of water in the preparation of the fermentation medium comprises a solution comprising an element selected from the group consisting of one or more of Zn, Co, Ni, one or more of W, Se. Blending with a solution comprising an element selected from the group consisting of an amount effective to provide a fermentation medium having a conductivity of about 30 mS / cm or less. This fermentation medium requires about 10% to about 40% less water than a fermentation medium with more than about 3 mM phosphate.
These and other aspects, features and advantages of some aspects of the process will become more apparent from the following drawings.
詳細な説明
以下の説明は、限定的意味に解釈すべきでなく、単に例示実施形態の一般原理を説明する目的で構成される。本発明の範囲は、特許請求の範囲を参照して決定すべきである。
一態様では、発酵槽への栄養素供給原料中の栄養レベルは、発酵槽内における酢酸生成微生物による各栄養素の消費%が本質的に等しくなるように最適化される。不運なことに、培地中の消費される栄養素の量と結果として生じる栄養素の残量とのアンバランスが伝導率上昇をもたらし、発酵性能が低下する。伝導率上昇を軽減するためには、大量の水が必要とされる。与えられる栄養素と消費される栄養素のバランスを慎重に取ると、水の利用と栄養素の利用が低減することになる。この態様では、栄養培地及び発酵プロセスは、90%以上の栄養素が利用され、別の態様では、少なくとも約95%以上の栄養素が利用されるように栄養素の利用を最適化する。
本明細書に記載の培地及び酢酸生成菌を含むバイオリアクター内で行なわるシンガス発酵は、シンガス中のCOのアルコールと他の生成物への変換をもたらすのに有効である。この態様では、生産性をgの全アルコール/(L・日)として表されるSTY(空時収量(space time yield))として表すことができる。この態様では、プロセスは、少なくとも約10g以上の全アルコール/(L・日)のSTY(空時収量)を与えるのに有効である。可能なSTY値としては約10gの全アルコール/(L・日)〜約200gの全アルコール/(L・日)、別の態様では、約10gの全アルコール/(L・日)〜約160gの全アルコール/(L・日)、別の態様では、約10gの全アルコール/(L・日)〜約120gの全アルコール/(L・日)、別の態様では、約10gの全アルコール/(L・日)〜約80gの全アルコール/(L・日)、別の態様では、約20gの全アルコール/(L・日)〜約140gの全アルコール/(L・日)、別の態様では、約20gの全アルコール/(L・日)〜約100gの全アルコール/(L・日)、別の態様では、約40gの全アルコール/(L・日)〜約140gの全アルコール/(L・日)、別の態様では、約40gの全アルコール/(L・日)〜約100gの全アルコール/(L・日)が挙げられる。
DETAILED DESCRIPTION The following description should not be construed in a limiting sense, but is merely configured to illustrate the general principles of exemplary embodiments. The scope of the invention should be determined with reference to the claims.
In one aspect, the nutrient level in the nutrient feed to the fermentor is optimized such that the% consumption of each nutrient by the acetic acid producing microorganism in the fermentor is essentially equal. Unfortunately, the imbalance between the amount of nutrients consumed in the medium and the resulting nutrient balance results in increased conductivity and reduced fermentation performance. A large amount of water is required to reduce the increase in conductivity. Careful balancing of the nutrients provided and consumed will reduce the use of water and nutrients. In this aspect, the nutrient medium and fermentation process optimizes nutrient utilization such that 90% or more nutrients are utilized, and in another aspect, at least about 95% or more nutrients are utilized.
Syngas fermentation performed in a bioreactor containing the media and acetic acid producing bacteria described herein is effective to effect the conversion of CO in syngas to alcohol and other products. In this embodiment, productivity can be expressed as STY (space time yield) expressed as total alcohol in g / (L · day). In this embodiment, the process is effective to provide a STY (space time yield) of at least about 10 g total alcohol / (L · day). Possible STY values are about 10 g total alcohol / (L.day) to about 200 g total alcohol / (L.day), in another embodiment about 10 g total alcohol / (L.day) to about 160 g. Total alcohol / (L.day), in another embodiment, from about 10 g total alcohol / (L.day) to about 120 g total alcohol / (L.day), in another embodiment, about 10 g total alcohol / ( L.day) to about 80 g total alcohol / (L.day), in another embodiment, about 20 g total alcohol / (L.day) to about 140 g total alcohol / (L.day), in another embodiment. About 20 g of total alcohol / (L.day) to about 100 g of total alcohol / (L.day); in another embodiment, about 40 g of total alcohol / (L.day) to about 140 g of total alcohol / (L • Day), in another embodiment, about 40 g total alcohol / (L · day) to about 100 g total alcohol / (L · day).
定義
特に指定のない限り、本開示のためにこの明細書全体を通じて使用する下記用語は下記定義どおりであり、下記定義の単数形又は複数形のどちらをも含み得る。
任意量を修飾する用語「約」は、実社会の条件内、例えば、実験室、パイロットプラント、又は生産施設内で遭遇する当該量の変動を指す。例えば、「約」で修飾されるときに混合物又は分量において用いられる成分又は測定値の量には、生産プラント又は実験室内の実験条件下で測定する際に典型的に払われる注意の変動又は程度が含まれる。例えば、「約」で修飾されるときの生成物の成分の量には、プラント又は実験室内の複数の実験のバッチ間の変動及び分析方法に固有の変動が含まれる。「約」で修飾されるか否かにかかわらず、量は当該量と等価な量を包含する。本明細書で言及され、「約」で修飾されたいずれの量も本開示では「約」で修飾されない量として利用することもできる。
用語「ガス状基質」は非限定的意味で使用され、この用語には1種以上のガスを含有するか又は1種以上のガスから誘導される基質が含まれる。
用語「シンガス」又は「合成ガス」は、各種量の一酸化炭素と水素を含有するガス混合物に与えられる名称である合成ガスを意味する。生産方法の例としては、水素を生産するための天然ガス又は炭化水素の水蒸気改質、石炭のガス化及びいくつかのタイプの廃棄物・トゥー・エネルギーのガス化施設内のガス化が挙げられる。この名称は、合成天然ガス(SNG)を作り出す際及びアンモニア又はメタノールを生産するための中間体としてのそれらの使用に由来する。シンガスは可燃性であり、他の化学薬品の生産のための燃料源又は中間体として用いられることが多い。
Definitions Unless otherwise specified, the following terms used throughout this specification for purposes of this disclosure are as defined below and may include either the singular or plural number of the following definitions.
The term “about” modifying an arbitrary amount refers to the variation in that amount encountered in real-world conditions, eg, in a laboratory, pilot plant, or production facility. For example, the amount of a component or measurement used in a mixture or quantity when modified with “about” includes the variation or degree of attention typically taken when measuring under experimental conditions in a production plant or laboratory. Is included. For example, the amount of product components when modified by “about” includes variations between batches of multiple experiments in the plant or laboratory and variations inherent in the analytical method. Whether or not modified with “about”, an amount includes an equivalent amount. Any amount referred to herein and modified by “about” may be utilized as an amount not modified by “about” in the present disclosure.
The term “gaseous substrate” is used in a non-limiting sense, and the term includes substrates that contain or are derived from one or more gases.
The term “syngas” or “syngas” means syngas, the name given to gas mixtures containing various amounts of carbon monoxide and hydrogen. Examples of production methods include steam reforming of natural gas or hydrocarbons to produce hydrogen, gasification of coal and gasification in some types of waste-to-energy gasification facilities. . This name comes from their use as intermediates in producing synthetic natural gas (SNG) and producing ammonia or methanol. Syngas is flammable and is often used as a fuel source or intermediate for the production of other chemicals.
用語「発酵槽」には、1つ以上の容器及び/若しくは塔又は配管から成る発酵装置が含まれ、連続撹拌槽型反応器(Continuous Stirred Tank Reactor)(CSTR)、固定化細胞反応器(Immobilized Cell Reactor)(ICR)、トリクルベッド反応器(Trickle Bed Reactor)(TBR)、移動床バイオフィルム反応器(Moving Bed Biofilm Reactor)(MBBR)、気泡塔、ガスリフト発酵槽、膜反応器、例えば中空繊維膜バイオリアクター(Hollow Fibre Membrane Bioreactor)(HFMBR)、静的ミキサー、又は気液接触に適した他の容器若しくは他の装置が挙げられる。
用語「発酵」、「発酵プロセス」又は「発酵反応」等はプロセスの成長期と生成物生合成期の両方を包含する意図である。一態様では、発酵は、COのアルコールへの変換を意味する。
用語「細胞密度」は、発酵ブロスの単位体積当たりの微生物細胞の質量、例えば、グラム/リットルを意味する。
発酵プロセスに関して使用するとき、用語「効率を高める」、「向上した効率」等には、発酵における微生物の成長速度、消費された基質(例えば一酸化炭素)の体積又は質量当たりの生成された所望生成物(例えばアルコール)の体積又は質量、所望生成物の生産速度又は生産レベル、及び生成された所望生成物の、発酵の他の副生物と比較した相対的比率の1つ以上を高めることが含まれる。
本明細書で使用する場合、「全アルコール」には、エタノール、ブタノール、プロパノール及びメタノールが含まれる。一態様では、全アルコールは、少なくとも約75質量パーセント以上のエタノール、別の態様では、約80質量パーセント以上のエタノール、別の態様では、約85質量パーセント以上のエタノール、別の態様では、約90質量パーセント以上のエタノール、別の態様では、約95質量パーセント以上のエタノールを含み得る。別の態様では、全アルコールは約25質量パーセント以下のブタノールを含み得る。
用語「比CO取込み」は、分での単位時間当たりの微生物細胞の単位質量(g)によって消費されたミリモルでのCOの量、すなわちミリモル/グラム/分を意味する。
The term “fermentor” includes a fermenter consisting of one or more vessels and / or towers or pipes, such as a Continuous Stirred Tank Reactor (CSTR), an Immobilized Cell Reactor (Immobilized). Cell Reactor (ICR), Trickle Bed Reactor (TBR), Moving Bed Biofilm Reactor (MBBR), bubble column, gas lift fermenter, membrane reactor, e.g. hollow fiber A membrane fiber reactor (Hollow Fiber Membrane Bioreactor) (HFMBR), a static mixer, or other vessel or other device suitable for gas-liquid contact.
The terms “fermentation”, “fermentation process” or “fermentation reaction” and the like are intended to encompass both the growth phase and the product biosynthesis phase of the process. In one aspect, fermentation refers to the conversion of CO to alcohol.
The term “cell density” means the mass of microbial cells per unit volume of fermentation broth, eg grams / liter.
When used in reference to a fermentation process, the terms “enhance efficiency”, “enhanced efficiency”, etc. include the growth rate of microorganisms in the fermentation, the desired generated per volume or mass of substrate consumed (eg, carbon monoxide). To increase one or more of the volume or mass of the product (e.g. alcohol), the production rate or level of the desired product, and the relative proportion of the desired product produced compared to other by-products of the fermentation. included.
As used herein, “total alcohol” includes ethanol, butanol, propanol and methanol. In one embodiment, the total alcohol is at least about 75 weight percent or more ethanol, in another embodiment, about 80 weight percent or more, in another embodiment, about 85 weight percent or more ethanol, in another embodiment, about 90 weight percent. It may comprise greater than or equal to weight percent ethanol, and in another embodiment greater than or equal to about 95 weight percent ethanol. In another embodiment, the total alcohol may contain up to about 25 weight percent butanol.
The term “specific CO uptake” means the amount of CO in millimoles consumed by unit mass (g) of microbial cells per unit time in minutes, ie millimoles / gram / minute.
CO含有基質
CO含有基質には、COを含むいずれのガスも含まれ得る。この態様では、CO含有ガスには、シンガス、産業ガス、及びその混合物が含まれ得る。
いずれの既知源からシンガスを供給してもよい。一態様では、炭素質材料のガス化からシンガスを調達することができる。ガス化は、酸素の制限された供給下でのバイオマスの部分燃焼を伴う。結果として生じるガスは主にCOとH2を含む。この態様では、シンガスは少なくとも約10モル%のCO、一態様では、少なくとも約20モル%のCO、一態様では、約10〜約100モル%のCO、別の態様では、約20〜約100モル%のCO、別の態様では、約30〜約90モル%のCO、別の態様では、約40〜約80モル%のCO、別の態様では、約50〜約70モル%のCOを含有するであろう。適切なガス化の方法及び装置のいくつかの例は、米国特許出願第61/516,667号、第61/516,704号及び第61/516,646号(全て2011年4月6日に出願された)、並びに米国特許出願第13/427,144号、第13/427,193号及び第13/427,247号(全て2012年3月22日に出願された)に提供されており、これら全ての内容を参照によってここに援用する。
別の態様では、本プロセスは、高体積CO含有産業煙道ガスのようなガス状基質からのアルコールの生産を支援する適用性を有する。一部の態様では、COを含むガスは、炭素含有廃棄物、例えば、産業廃ガス又は他の廃棄物のガス化から誘導される。そのようなものとして、本プロセスは、そうでなければ環境に排出されるであろう炭素を捕獲するための有効なプロセスを提起する。産業煙道ガスの例として、鉄類製品製造、非鉄製品製造、石油精製プロセス、石炭のガス化、バイオマスのガス化、電力生産、カーボンブラック生産、アンモニア生産、メタノール生産及びコークス製造中に生成されるガスが挙げられる。
CO-containing substrate
The CO-containing substrate can include any gas containing CO. In this aspect, the CO-containing gas can include syngas, industrial gas, and mixtures thereof.
The syngas may be supplied from any known source. In one aspect, syngas can be procured from the gasification of the carbonaceous material. Gasification involves the partial combustion of biomass under a limited supply of oxygen. The resulting gas mainly comprising CO and H 2. In this embodiment, the syngas is at least about 10 mol% CO, in one embodiment at least about 20 mol% CO, in one embodiment from about 10 to about 100 mol% CO, in another embodiment from about 20 to about 100. Mol% CO, in another embodiment about 30 to about 90 mol% CO, in another embodiment about 40 to about 80 mol% CO, in another embodiment about 50 to about 70 mol% CO. Will contain. Some examples of suitable gasification methods and apparatus include U.S. Patent Application Nos. 61 / 516,667, 61 / 516,704 and 61 / 516,646 (all filed April 6, 2011), and Provided in U.S. Patent Applications Nos. 13 / 427,144, 13 / 427,193, and 13 / 427,247 (all filed March 22, 2012), the entire contents of which are hereby incorporated by reference. .
In another aspect, the process has applicability to support the production of alcohol from gaseous substrates such as high volume CO containing industrial flue gases. In some aspects, the CO-containing gas is derived from the gasification of carbon-containing waste, such as industrial waste gas or other waste. As such, the process presents an effective process for capturing carbon that would otherwise be emitted to the environment. Examples of industrial flue gas produced during ferrous product manufacturing, non-ferrous product manufacturing, petroleum refining process, coal gasification, biomass gasification, power production, carbon black production, ammonia production, methanol production and coke production Gas.
CO含有基質の組成に応じて、CO含有基質を発酵プロセスに直接与えるか又は適切なH2対COモル比を含むようにさらに改変してよい。一態様では、発酵槽に与えるCO含有基質は、約0.2以上のH2対COモル比、別の態様では、約0.25以上のH2対COモル比、及び別の態様では、約0.5以上のH2対COモル比を有する。別の態様では、発酵槽に与えられるCO含有基質は約40モルパーセント以上のCOプラスH2及び約30モルパーセント以下のCO、別の態様では、約50モルパーセント以上のCOプラスH2及び約35モルパーセント以下のCO、別の態様では、約80モルパーセント以上のCOプラスH2及び約20モルパーセント以下のCOを含み得る。
一態様では、CO含有基質は主にCOとH2を含む。この態様では、CO含有基質は、少なくとも約10モル%のCO、一態様では、少なくとも約20モル%、一態様では、約10〜約100モル%、別の態様では、約20〜約100モル%のCO、別の態様では、約30〜約90モル%のCO、別の態様では、約40〜約80モル%のCO、別の態様では、約50〜約70モル%のCOを含有するであろう。CO含有基質は、少なくとも約0.75のCO/CO2比、別の態様では、少なくとも約1.0、別の態様では、少なくとも約1.5のCO/CO2比を有するであろう。
Depending on the composition of the CO-containing substrate, the CO-containing substrate may be provided directly to the fermentation process or further modified to include an appropriate H 2 to CO molar ratio. In one aspect, the CO-containing substrate provided to the fermentor has an H 2 to CO molar ratio of about 0.2 or higher, in another aspect, an H 2 to CO molar ratio of about 0.25 or higher, and in another aspect, about 0.5 or higher. H 2 to CO molar ratio. In another embodiment, the CO-containing substrate provided to the fermentor is about 40 mole percent or more CO plus H 2 and about 30 mole percent or less CO, in another embodiment about 50 mole percent or more CO plus H 2 and about Up to about 35 mole percent CO, and in another embodiment, about 80 mole percent or more CO plus H 2 and about 20 mole percent or less CO.
In one embodiment, CO-containing substrate mainly comprising CO and H 2. In this embodiment, the CO-containing substrate is at least about 10 mol% CO, in one embodiment at least about 20 mol%, in one embodiment from about 10 to about 100 mol%, in another embodiment from about 20 to about 100 mol%. % CO, in another embodiment about 30 to about 90 mol% CO, in another embodiment about 40 to about 80 mol% CO, in another embodiment about 50 to about 70 mol% CO Will do. CO-containing substrate is at least about 0.75 of CO / CO 2 ratio, in another aspect, at least about 1.0, in another aspect, it will have at least about 1.5 CO / CO 2 ratio.
一態様では、ガス分離器を配置して、ガス流の少なくとも一部を実質的に分離する。この部分には1種以上の成分が含まれる。例えば、ガス分離器は、下記成分:CO、CO2、H2を含むガス流からCO2を分離し、CO2をCO2除去器に通し、ガス流の残り(CO及びH2を含む)をバイオリアクターに通すことができる。技術上周知のいずれのガス分離器をも利用し得る。この態様では、発酵槽に与えられるシンガスは約10モル%以下のCO2、別の態様では、約1モル%以下のCO2、別の態様では、約0.1モル%以下のCO2を有するであろう。
ある一定のガス流は高濃度のCOと低濃度のH2を含んでよい。一態様では、より高いアルコール生産効率及び/又は全体的な炭素捕獲を達成するため、基質流の組成を最適化するのが望ましいことがある。例えば、基質流をバイオリアクターに通す前に基質流中のH2濃度を高めてよい。
本発明の特定の態様によれば、2種以上の起源からの流れを組み合わせ及び/又はブレンドして所望の及び/又は最適化基質流を生成することができる。例えば、製鋼所転炉からの排ガス等の高濃度COを含む流れを製鋼所コークス炉からのオフガス等の高濃度H2を含む流れと組み合わせることができる。
ガス状CO含有基質の組成によっては、CO含有基質を発酵に導入する前に処理して、粉塵粒子等のいずれの望ましくない不純物をも除去するのが望ましいこともある。例えば、既知の方法を用いてガス状基質をろ過又は洗浄することができる。
In one aspect, a gas separator is disposed to substantially separate at least a portion of the gas stream. This part contains one or more components. For example, a gas separator, the following ingredients: CO, CO 2 from a gas stream containing CO 2, H 2 was separated, passed through a CO 2 to CO 2 remover (including CO and H 2) remaining gas stream Can be passed through a bioreactor. Any gas separator known in the art can be utilized. In this embodiment, syngas given fermenter is about 10 mole percent of CO 2, in another embodiment, from about 1 mole percent of CO 2, in another embodiment, have from about 0.1 mol% or less of CO 2 I will.
Certain gas stream may comprise of H 2 of a high concentration of CO and a low concentration. In one aspect, it may be desirable to optimize the composition of the substrate stream to achieve higher alcohol production efficiency and / or overall carbon capture. For example, the H 2 concentration in the substrate stream may be increased before passing the substrate stream through the bioreactor.
According to certain aspects of the invention, streams from two or more sources can be combined and / or blended to produce a desired and / or optimized substrate stream. For example, a flow containing high concentration CO such as exhaust gas from a steel mill converter can be combined with a flow containing high concentration H 2 such as off-gas from a steel mill coke oven.
Depending on the composition of the gaseous CO-containing substrate, it may be desirable to treat the CO-containing substrate prior to introduction into the fermentation to remove any undesirable impurities such as dust particles. For example, the gaseous substrate can be filtered or washed using known methods.
バイオリアクターの設計と操作
発酵槽設計の詳細は米国特許出願第13/471,827号及び第13/471,858号(両方とも2012年5月15日に出願された)、並びに2012年5月16日に出願された米国特許出願第13/473,167号に記載されており、これら全ての内容を参照によってここに援用する。
一態様によれば、培地を反応器に添加して発酵プロセスを開始する。培地組成物のいくつかの例は、2012年5月22日に出願された米国特許出願第61/650,098号及び第61/650,093号、並びに2001年7月23日に出願された米国特許第7,285,402号に記載されており、これら全ての内容を参照によってここに援用する。培地を滅菌して望ましくない微生物を除去することができ、所望の微生物を反応器に接種する。滅菌は常に必要なわけではない。
一態様では、利用微生物には酢酸生成菌が含まれる。有用な酢酸生成菌の例としては、クロストリジウム属のもの、例えばクロストリジウム・リュングダリイの株(WO 2000/68407、EP 117309、米国特許第5,173,429号、第5,593,886号及び第6,368,819号、WO 1998/00558及びWO 2002/08438に記載されたものを含めて)、クロストリジウム・オートエタノゲナム(Clostridium autoethanogenum)の株(DSMZ, GermanyのDSM 10061及びDSM 19630)(WO 2007/117157及びWO 2009/151342に記載されたものを含めて)及びクロストリジウム・ラグスダレイ(Clostridium ragsdalei)(P11, ATCC BAA-622)及びアルカリバクルム・バッキ(Alkalibaculum bacchi)(CP11, ATCC BAA-1772)(それぞれ米国特許第7,704,723号及び2010年4月29日にOklahoma EPSCoR Annual State Conferenceで提出された“Biofuels and Bioproducts from Biomass-Generated Synthesis Gas”,Hasan Atiyehに記載されたものを含めて)及び米国特許出願第2007/0276447号に記載のクロストリジウム・カルボキシディボランス(Clostridium carboxidivorans)(ATCC PTA-7827)が挙げられる。他の適切な微生物には、ムーレラ属(Moorella)のもの(ムーレラ種HUC22-1を含めて)、及びカーボキシドサーマス(Carboxydothermus)属のものがある。これらの各参考文献を参照によってここに援用する。2種以上の微生物の混合培養を使用してよい。
Bioreactor design and operation Fermenter design details can be found in U.S. Patent Application Nos. 13 / 471,827 and 13 / 471,858 (both filed on May 15, 2012) and filed on May 16, 2012. No. 13 / 473,167, the entire contents of which are hereby incorporated by reference.
According to one aspect, media is added to the reactor to initiate the fermentation process. Some examples of media compositions include U.S. Patent Applications 61 / 650,098 and 61 / 650,093 filed May 22, 2012, and U.S. Patent No. 7,285,402 filed July 23, 2001. All of which are hereby incorporated by reference. The medium can be sterilized to remove unwanted microorganisms and the desired microorganisms are inoculated into the reactor. Sterilization is not always necessary.
In one aspect, the microorganisms utilized include acetic acid producing bacteria. Examples of useful acetic acid producing bacteria include those of the genus Clostridium, such as the strains of Clostridium Lungdalii (WO 2000/68407, EP 117309, US Pat. Nos. 5,173,429, 5,593,886 and 6,368,819, WO 1998/00558 and WO Strains of Clostridium autoethanogenum (including those described in 2002/08438) (DSMZ, DSM 10061 and DSM 19630 in Germany) (described in WO 2007/117157 and WO 2009/151342) And Clostridium ragsdalei (P11, ATCC BAA-622) and Alkalibaculum bacchi (CP11, ATCC BAA-1772) (US Pat. Nos. 7,704,723 and April 2010, respectively) Described in “Biofuels and Bioproducts from Biomass-Generated Synthesis Gas” filed at Oklahoma EPSCoR Annual State Conference on 29th) and US Patent Application No. 2007/0276447 Clostridium carboxidivorans (ATCC PTA-7827). Other suitable microorganisms include those of the genus Moorella (including Mourela sp. HUC22-1) and those of the genus Carboxydothermus. Each of these references is hereby incorporated by reference. A mixed culture of two or more microorganisms may be used.
有用な細菌のいくつかの例として、アセトゲニウム・キブイ(Acetogenium kivui)、アセトバクテリウム・ノテラエ(Acetoanaerobium noterae)、アセトバクテリウム・ウッディイ(Acetobacterium woodii)、アルカリバクルム・バッキ(Alkalibaculum bacchi)CP11(ATCC BAA-1772)、ブラウティア・プロダクタ(Blautia producta)、ブチリバクテリウム・メチロトロフィカム(Butyribacterium methylotrophicum)、カルダナエロバクター・サブテラネウス(Caldanaerobacter subterraneous)、カルダナエロバクター・サブテラネウス・パシフィカム(Caldanaerobacter subterraneous pacificus)、カーボキシドサーマス・ハイドロゲノフォルマンス(Carboxydothermus hydrogenoformans)、クロストリジウム・アセチカム(Clostridium aceticum)、クロストリジウム・アセトブチリカム(Clostridium acetobutylicum)、クロストリジウム・アセトブチリカムP262(DSMZ GermanyのDSM 19630)、クロストリジウム・オートエタノゲナム(DSMZ GermanyのDSM 19630)、クロストリジウム・オートエタノゲナム(DSMZ GermanyのDSM 10061)、クロストリジウム・オートエタノゲナム(DSMZ GermanyのDSM 23693)、クロストリジウム・オートエタノゲナム(DSMZ GermanyのDSM 24138)、クロストリジウム・カルボキシディボランスP7(ATCC PTA-7827)、クロストリジウム・コスカチイ(Clostridium coskatii)(ATCC PTA-10522)、クロストリジウム・ドラケイ(Clostridium drakei)、クロストリジウム・リュングダリイPETC(ATCC 49587)、クロストリジウム・リュングダリイERI2(ATCC 55380)、クロストリジウム・リュングダリイC-01(ATCC 55988)、クロストリジウム・リュングダリイO-52(ATCC 55889)、クロストリジウム・マグナム(Clostridium magnum)、クロストリジウム・パストゥリアヌム(Clostridium pasteurianum)(DSMZ GermanyのDSM 525)、クロストリジウム・ラグスダリ(Clostridium ragsdali)P11(ATCC BAA-622)、クロストリジウム・スカトロゲネス(Clostridium scatologenes)、クロストリジウム・サーモアセチカム(Clostridium thermoaceticum)、クロストリジウム・ウルツネンセ(Clostridium ultunense)、デスルホトマクルム・クズネツォビイ(Desulfotomaculum kuznetsovii)、ユーバクテリウム・リモスム(Eubacterium limosum)、ゲオバクター・スルフレデュセンス(Geobacter sulfurreducens)、メタノサルシナ・アセチボランス(Methanosarcina acetivorans)、メタノサルシナ・バーケリ(Methanosarcina barkeri)、モーレラ・サーモアセティカ(Morrella thermoacetica)、モーレラ・サーモオートトロフィカ(Morrella thermoautotrophica)、オキソバクター・フェニジイ(Oxobacter pfennigii)、ペプトストレプトコッカス・プロダクツス(Peptostreptococcus productus)、ルミノコッカス・プロダクツス(Ruminococcus productus)、サーモアネロバクター・キブイ(Thermoanaerobacter kivui)、及びその混合物が挙げられる。 Some examples of useful bacteria include Acetogenium kivui, Acetoanaerobium noterae, Acetobacterium woodii, Alkalibaculum bacchi CP11 (ATCC) BAA-1772), Blautia producta, Butyribacterium methylotrophicum, Caldanaerobacter subterraneous, Caldanaerobacter subterraneus Pacifica ), Carboxydothermus hydrogenoformans, Clostridium aceticum, Clostridium acetobutylicum, Clostridium acetobutylicum P262 (DSMZ) DSM 19630 from Germany, Clostridium autoethanogenum (DSM 19630 from DSMZ Germany), Clostridium autoethanogenum (DSM 10061 from DSMZ Germany), Clostridium autoethanogenum (DSM 23693 from DSMZ Germany) (DSMZ 138, DSMZ Germany), Clostridium carboxydivorance P7 (ATCC PTA-7827), Clostridium coskatii (ATCC PTA-10522), Clostridium drakei, Clostridium lyngdalii PETC (ATCC 49587) ), Clostridium Lungdalii ERI2 (ATCC 55380), Clostridium Lungdalii C-01 (ATCC 55988), Clostridium Lungdalii O-52 (ATCC 55889), Clostridium magnum, Clostridium pasteurianum (D) German y DSM 525), Clostridium ragsdali P11 (ATCC BAA-622), Clostridium scatologenes, Clostridium thermoaceticum, Clostridium ultunense, Clostridium desulfun ultunense Desulfotomaculum kuznetsovii, Eubacterium limosum, Geobacter sulfurreducens, Methanosarcina acetivorans, Methanosarcina acetivorans, Methanosarcina barkeres Morrella thermoacetica, Morrella thermoautotrophica, Oxobacter pfennigii, Peptostreptococcus productus, Lumino Listed are Coccus products (Ruminococcus productus), Thermoanaerobacter kivui, and mixtures thereof.
望ましくは、所望の発酵(例えばCO→エタノール)が起こるのに適した条件下で発酵を行なうべきである。考慮すべき反応条件としては、圧力、温度、ガス流速、液体流速、培地のpH、培地の酸化還元電位、撹拌速度(連続撹拌槽型反応器を使用する場合)、接種レベル、液相内のCOが律速にならないことを確実にするための最大ガス基質濃度、及び生成物抑制を回避するための最大生成物濃度が挙げられる。
本発明の方法を用いて、COの溶液中への移動速度が培養の取込み速度未満であるようにCOが限られている微生物培養の生存能を持続することができる。このような状況は、COを含む基質を微生物培養に連続的に供給せず;物質移動速度が低く;又は不十分なCOが基質流中に存在するときに生じて、最適温度で培養の生命力を持続し得る。該実施形態では、微生物培養は、液体栄養培地に溶解したCOを急速に枯渇させ、さらなる基質は十分速くは与えられないので、限られた基質になるであろう。
Desirably, the fermentation should be performed under conditions suitable for the desired fermentation (eg, CO → ethanol) to occur. Reaction conditions to consider include pressure, temperature, gas flow rate, liquid flow rate, medium pH, medium redox potential, agitation rate (when using a continuous stirred tank reactor), inoculation level, liquid phase A maximum gas substrate concentration to ensure that CO is not rate limiting and a maximum product concentration to avoid product suppression.
The method of the present invention can be used to sustain the viability of microbial cultures with limited CO such that the rate of transfer of CO into the solution is less than the rate of uptake of the culture. This situation occurs when the substrate containing CO is not continuously fed to the microbial culture; the mass transfer rate is low; or when insufficient CO is present in the substrate stream and the vitality of the culture at the optimum temperature. Can last. In such embodiments, the microbial culture will rapidly deplete CO dissolved in the liquid nutrient medium, and no additional substrate will be provided fast enough, so it will be a limited substrate.
スタートアップ:接種したら、微生物の初期集団を供給するのに有効な初期供給ガスの供給速度を確立する。流出ガスを分析して流出ガスの内容を判定する。ガス分析の結果を用いて供給ガス速度を制御する。この態様では、プロセスは、約0.5〜約0.9、別の態様では、約0.6〜約0.8、別の態様では、約0.5〜約0.7、別の態様では、約0.5〜約0.6の計算CO濃度対初期細胞密度比を与える。
別の態様では、発酵プロセスは、約0.15mM〜約0.70mM、別の態様では、約0.15mM〜約0.50mM、別の態様では、約0.15mM〜約0.35mM、別の態様では、約0.20mM〜約0.30mM、別の態様では、約0.23mM〜約0.27mMの、発酵培地内の初期計算CO濃度を与えるのに有効な量でシンガスを発酵培地に与える工程を含む。このプロセスは、開始細胞密度に比べて細胞密度を増やすのに有効である。
Startup : Once inoculated, establish an initial feed gas supply rate effective to supply an initial population of microorganisms. The effluent gas is analyzed to determine the content of the effluent gas. The gas analysis results are used to control the feed gas rate. In this aspect, the process is performed at a calculated CO concentration pair of about 0.5 to about 0.9, in another aspect about 0.6 to about 0.8, in another aspect about 0.5 to about 0.7, and in another aspect about 0.5 to about 0.6. Gives the initial cell density ratio.
In another aspect, the fermentation process is from about 0.15 mM to about 0.70 mM, in another aspect, from about 0.15 mM to about 0.50 mM, in another aspect, from about 0.15 mM to about 0.35 mM, in another aspect, from about 0.20. providing syngas to the fermentation medium in an amount effective to provide an initial calculated CO concentration in the fermentation medium of from mM to about 0.30 mM, and in another embodiment from about 0.23 mM to about 0.27 mM. This process is effective to increase the cell density compared to the starting cell density.
スタートアップ後:所望レベルに達したら、液相及び細胞物質を反応器から取り出して培地を補充する。このプロセスは、細胞密度を約2.0グラム/リットル以上、別の態様では、約2〜約30グラム/リットル、別の態様では、約2〜約25グラム/リットル、別の態様では、約2〜約20グラム/リットル、別の態様では、約2〜約10グラム/リットル、別の態様では、約2〜約8グラム/リットル、別の態様では、約3〜約30グラム/リットル、別の態様では、約3〜約6グラム/リットル、別の態様では、約4〜約5グラム/リットルに増やすのに有効である。
一態様では、本プロセスは、Zn(プア培地とも呼ばれる)、Co、Niの1種以上からから成る群より選択される元素を含む第1溶液を、W及びSeの1種以上から成る群より選択される元素を含む第2溶液と、約30mS/cm以下の伝導率を有する発酵培地を与えるのに有効な量でブレンドする工程を含むプロセスにより与えられる発酵培地を含む。別の態様では、発酵培地は、約1〜約30mS/cm、別の態様では、約1〜約25mS/cm、別の態様では、約1〜約20mS/cm、別の態様では、約1〜約15mS/cm、別の態様では、約1〜約10mS/cm、別の態様では、約1〜約5mS/cm、別の態様では、約1〜約4mS/cm、別の態様では、約1〜約3mS/cm、別の態様では、約1〜約2mS/cm、別の態様では、約2〜約30mS/cm、別の態様では、約2〜約25mS/cm、別の態様では、約2〜約20mS/cm、別の態様では、約2〜約15mS/cm、別の態様では、約2〜約10mS/cm、別の態様では、約2〜約5mS/cm、約2〜約4mS/cm、別の態様では、約2〜約3mS/cm、別の態様では、約3〜約30mS/cm、別の態様では、約3〜約25mS/cm、別の態様では、約3〜約20mS/cm、別の態様では、約3〜約15mS/cm、別の態様では、約3〜約10mS/cm、別の態様では、約3〜約5mS/cm、別の態様では、約4〜約30mS/cm、別の態様では、約4〜約25mS/cm、別の態様では、約4〜約20mS/cm、別の態様では、約4〜約15mS/cm、別の態様では、約4〜約10mS/cm、別の態様では、約4〜約5mS/cmの伝導率を有する。
別の態様では、元素のブレンドは、580nmで約0.70以下の光学密度を有する。別の態様では、ブレンドは、約0〜約0.70、別の態様では、約0.001〜約0.65、別の態様では、約0.01〜約0.65、別の態様では、約0.01〜約0.50、別の態様では、約0.01〜約0.45の光学密度を有する。この態様では、任意の既知の方法によって濁度を決定可能である。光学密度測定のいくつかの例は、参照によってその内容全体をここに援用するEPA Guidance Manual, Turbidity Processes, April 1999に記載されている。
別の態様では、発酵培地は約14mM未満のホスファートを有する。関連態様では、発酵培地は約2〜約14mMのホスファート、別の態様では、約3〜約12mMのホスファート、別の態様では、約3〜約6mMのホスファート、別の態様では、約1〜約3mMのホスファート、別の態様では、約1〜約2mMのホスファート、別の態様では、約2〜約3mMのホスファートを有する。
After start-up : When the desired level is reached, remove the liquid phase and cellular material from the reactor and replenish the medium. This process can have a cell density of about 2.0 grams / liter or more, in another embodiment about 2 to about 30 grams / liter, in another embodiment about 2 to about 25 grams / liter, in another embodiment about 2 to about 30 grams / liter. About 20 grams / liter, in another embodiment about 2 to about 10 grams / liter, in another embodiment about 2 to about 8 grams / liter, in another embodiment about 3 to about 30 grams / liter, another In embodiments, it is effective to increase to about 3 to about 6 grams / liter, and in another embodiment about 4 to about 5 grams / liter.
In one aspect, the process includes a first solution comprising an element selected from the group consisting of one or more of Zn (also referred to as poor medium), Co, and Ni, from the group consisting of one or more of W and Se. A second solution containing a selected element and a fermentation medium provided by a process comprising a step of blending in an amount effective to provide a fermentation medium having a conductivity of about 30 mS / cm or less. In another embodiment, the fermentation medium is about 1 to about 30 mS / cm, in another embodiment about 1 to about 25 mS / cm, in another embodiment about 1 to about 20 mS / cm, in another embodiment about 1 To about 15 mS / cm, in another embodiment from about 1 to about 10 mS / cm, in another embodiment from about 1 to about 5 mS / cm, in another embodiment from about 1 to about 4 mS / cm, in another embodiment, About 1 to about 3 mS / cm, in another embodiment about 1 to about 2 mS / cm, in another embodiment about 2 to about 30 mS / cm, in another embodiment about 2 to about 25 mS / cm, another embodiment About 2 to about 20 mS / cm, in another embodiment about 2 to about 15 mS / cm, in another embodiment about 2 to about 10 mS / cm, in another embodiment about 2 to about 5 mS / cm, about 2 to about 4 mS / cm, in another embodiment about 2 to about 3 mS / cm, in another embodiment about 3 to about 30 mS / cm, in another embodiment about 3 to about 25 mS / cm, in another embodiment About 3 to about 20 mS / cm, in another embodiment about 3 to about 15 mS / cm, in another embodiment about 3 to about 10 mS / cm, in another embodiment about 3 to about 5 mS / cm, another In embodiments, about 4 to about 30 mS / cm, in another embodiment, about 4 About 25 mS / cm, in another embodiment about 4 to about 20 mS / cm, in another embodiment about 4 to about 15 mS / cm, in another embodiment about 4 to about 10 mS / cm, in another embodiment about It has a conductivity of 4 to about 5 mS / cm.
In another embodiment, the blend of elements has an optical density of about 0.70 or less at 580 nm. In another aspect, the blend is from about 0 to about 0.70, in another aspect from about 0.001 to about 0.65, in another aspect, from about 0.01 to about 0.65, in another aspect, from about 0.01 to about 0.50, in another aspect. Has an optical density of about 0.01 to about 0.45. In this embodiment, turbidity can be determined by any known method. Some examples of optical density measurements are described in the EPA Guidance Manual, Turbidity Processes, April 1999, the entire contents of which are incorporated herein by reference.
In another embodiment, the fermentation medium has less than about 14 mM phosphate. In a related embodiment, the fermentation medium is about 2 to about 14 mM phosphate, in another embodiment about 3 to about 12 mM phosphate, in another embodiment about 3 to about 6 mM phosphate, in another embodiment about 1 to about 3 mM phosphate, in another embodiment from about 1 to about 2 mM phosphate, in another embodiment from about 2 to about 3 mM phosphate.
一態様では、本プロセスは、エタノール1米国ガロン(3.8リットル)当たり発酵培地に供給される約2米国ガロン(7.6リットル)以下の水を利用するのに有効である。別の態様では、本プロセスは、エタノール1ガロン当たり約0.5〜約2ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約1.8ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約1.5ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約1.35ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約1.2ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約1ガロンの水、別の態様では、エタノール1ガロン当たり約0.5〜約0.9ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約2ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約1.75ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約1.5ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約1.35ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約1.2ガロンの水、別の態様では、エタノール1ガロン当たり約0.75〜約1ガロンの水、別の態様では、エタノール1ガロン当たり約1〜約2ガロンの水、別の態様では、エタノール1ガロン当たり約1〜約1.75ガロンの水、別の態様では、エタノール1ガロン当たり約1〜約1.5ガロンの水、別の態様では、エタノール1ガロン当たり約1〜約1.35ガロンの水、別の態様では、エタノール1ガロン当たり約1〜約1.2ガロンの水、別の態様では、エタノール1ガロン当たり約1.5〜約2ガロンの水、別の態様では、エタノール1ガロン当たり約1.5〜約1.75ガロンの水、別の態様では、エタノール1ガロン当たり約1.75〜約2ガロンの水を利用するのに有効である。
別の態様では、発酵培地は、約3mM以上のホスファートを有する発酵培地より約10%〜約40%少ない水を必要とする。別の態様では、発酵培地は、約3mM以上のホスファートを有する発酵培地より約10%〜約30%少ない水、別の態様では、約10%〜約20%少ない水、別の態様では、約15%〜約40%少ない水、別の態様では、約15%〜約30%少ない水、別の態様では、約15%〜約20%少ない水、別の態様では、約20%〜約40%少ない水、別の態様では、約20%〜約30%少ない水、別の態様では、約25%〜約30%少ない水を必要とする。別の態様では、ホスファート濃度は、約2〜約2.5mM、別の態様では、約2.5mM〜約3.0mMであってよく、かつ指示範囲の水の低減を得るのに有効であり得る。
In one aspect, the process is effective to utilize no more than about 2 US gallons (7.6 liters) of water supplied to the fermentation medium per 1 US gallon (3.8 liters) of ethanol. In another embodiment, the process comprises from about 0.5 to about 2 gallons of water per gallon of ethanol, in another embodiment, from about 0.5 to about 1.8 gallons of water per gallon of ethanol, in another embodiment, from about 0.5 to about 1 gallon of ethanol. 0.5 to about 1.5 gallons of water, in another embodiment about 0.5 to about 1.35 gallons of water per gallon of ethanol, in another embodiment, about 0.5 to about 1.2 gallons of water per gallon of ethanol, in another embodiment, ethanol About 0.5 to about 1 gallon of water per gallon, in another embodiment about 0.5 to about 0.9 gallon of water per gallon of ethanol, in another embodiment about 0.75 to about 2 gallons of water per gallon of ethanol, another In embodiments, from about 0.75 to about 1.75 gallons of water per gallon of ethanol, in another embodiment, from about 0.75 to about 1.5 gallons of water per gallon of ethanol, in another embodiment, from about 0.75 to about 1.35 gallons per gallon of ethanol. Water, another state From about 0.75 to about 1.2 gallons of water per gallon of ethanol; in another embodiment, from about 0.75 to about 1 gallon of water per gallon of ethanol; in another embodiment, from about 1 to about 2 gallons of water per gallon of ethanol In another embodiment, from about 1 to about 1.75 gallons of water per gallon of ethanol; in another embodiment, from about 1 to about 1.5 gallons of water per gallon of ethanol; in another embodiment, from about 1 to about 1 per gallon of ethanol 1.35 gallons of water, in another embodiment from about 1 to about 1.2 gallons of water per gallon of ethanol, in another embodiment, from about 1.5 to about 2 gallons of water per gallon of ethanol, in another embodiment, per gallon of ethanol Effective from about 1.5 to about 1.75 gallons of water, in another embodiment, from about 1.75 to about 2 gallons of water per gallon of ethanol.
In another embodiment, the fermentation medium requires about 10% to about 40% less water than a fermentation medium with about 3 mM or more phosphate. In another aspect, the fermentation medium is about 10% to about 30% less water, in another aspect about 10% to about 20% less water, in another aspect about about 3% or more phosphate medium. 15% to about 40% less water, in another embodiment about 15% to about 30% less water, in another embodiment about 15% to about 20% less water, in another embodiment, about 20% to about 40 % Water, in another embodiment, about 20% to about 30% less water, in another embodiment, about 25% to about 30% less water. In another embodiment, the phosphate concentration may be from about 2 to about 2.5 mM, in another embodiment from about 2.5 mM to about 3.0 mM, and may be effective to obtain the indicated range of water reduction.
別の態様では、発酵培地は、細胞1グラム当たり毎分約0.005μg以上のZn、細胞1グラム当たり毎分約0.0002μg以上のCo、細胞1グラム当たり毎分約0.003μg以上のNi、細胞1グラム当たり毎分約0.039μg以上のW、及び細胞1グラム当たり毎分約0.001μg以上のSeが与えられる。この態様では、発酵培地は、下記量の下記元素の1種以上を含み得る:
Zn:一態様では、細胞1グラム当たり毎分約0.005〜約0.11μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.09μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.065μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.04μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.075μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.055μg、別の態様では、細胞1グラム当たり毎分約0.02〜約0.075μg、別の態様では、細胞1グラム当たり毎分約0.02〜約0.055μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.04μgのZn供給速度を必要とするであろう;
Co:一態様では、細胞1グラム当たり毎分約0.002〜約0.05μg、別の態様では、細胞1グラム当たり毎分約0.002〜約0.04μg、別の態様では、細胞1グラム当たり毎分約0.002〜約0.03μg、別の態様では、細胞1グラム当たり毎分約0.002〜約0.02μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.035μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.025μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.035μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.025μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.018μgのCo供給速度を必要とするであろう;
Ni:一態様では、細胞1グラム当たり毎分約0.003〜約0.055μg、別の態様では、細胞1グラム当たり毎分約0.003〜約0.045μg、別の態様では、細胞1グラム当たり毎分約0.003〜約0.035μg、別の態様では、細胞1グラム当たり毎分約0.003〜約0.02μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.04μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.03μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.04μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.03μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.02μgのNi供給速度を必要とするであろう;
W:一態様では、細胞1グラム当たり毎分約0.035〜約0.80μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.65μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.47μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.30μg、別の態様では、細胞1グラム当たり毎分約0.075〜約0.55μg、別の態様では、細胞1グラム当たり毎分約0.075〜約0.40μg、別の態様では、細胞1グラム当たり毎分約0.155〜約0.55μg、別の態様では、細胞1グラム当たり毎分約0.155〜約0.40μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.29μgのW供給速度を必要とするであろう;
Se:一態様では、細胞1グラム当たり毎分約0.001〜約0.03μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.65μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.47μg、別の態様では、細胞1グラム当たり毎分約0.035〜約0.30μg、別の態様では、細胞1グラム当たり毎分約0.075〜約0.55μg、別の態様では、細胞1グラム当たり毎分約0.075〜約0.40μg、別の態様では、細胞1グラム当たり毎分約0.155〜約0.55μg、別の態様では、細胞1グラム当たり毎分約0.155〜約0.40μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.01μgのSe供給速度を必要とするであろう。
In another embodiment, the fermentation medium comprises about 0.005 μg or more of Zn per gram of cells, about 0.0002 μg or more of Co per gram of cells, about 0.003 μg or more of Ni per gram of cells, 1 cell More than about 0.039 μg W per gram per minute and more than about 0.001 μg Se per gram per cell are given. In this aspect, the fermentation medium can include one or more of the following elements in the following amounts:
Zn : In one aspect, from about 0.005 to about 0.11 μg per gram of cells per gram, in another aspect, from about 0.005 to about 0.09 μg per gram of cells, in another aspect, about 0.005 per gram of cells per minute. To about 0.065 μg, in another embodiment, from about 0.005 to about 0.04 μg per gram of cells per gram, in another embodiment, from about 0.01 to about 0.075 μg per gram of cells, in another embodiment, per gram of cells. From about 0.01 to about 0.055 μg per minute, in another embodiment, from about 0.02 to about 0.075 μg per gram of cells, in another embodiment, from about 0.02 to about 0.055 μg per gram of cells; A specific ethanol productivity of L / day / gram (cells) would require a Zn feed rate of about 0.04 μg per minute per gram of cells;
Co : In one aspect, from about 0.002 to about 0.05 μg per gram of cells per gram, in another aspect, from about 0.002 to about 0.04 μg per gram of cells, in another aspect, about 0.002 per minute per gram of cells. To about 0.03 μg, in another embodiment, from about 0.002 to about 0.02 μg per gram of cells per gram, in another embodiment, from about 0.005 to about 0.035 μg per gram of cells, in another embodiment, per gram of cells From about 0.005 to about 0.025 μg per minute, in another embodiment, from about 0.01 to about 0.035 μg per gram of cells, in another embodiment, from about 0.01 to about 0.025 μg per gram of cells; A specific ethanol productivity of L / day / gram (cells) would require a Co feed rate of about 0.018 μg per minute per gram of cells;
Ni : In one aspect, from about 0.003 to about 0.055 μg per gram of cells per gram, in another aspect, from about 0.003 to about 0.045 μg per gram of cells, in another aspect, from about 0.003 per gram of cells per minute. To about 0.035 μg, in another embodiment, from about 0.003 to about 0.02 μg per gram of cells per gram, in another embodiment, from about 0.005 to about 0.04 μg per gram of cells, in another embodiment, per gram of cells. From about 0.005 to about 0.03 μg per minute, in another embodiment, from about 0.01 to about 0.04 μg per gram of cell, in another embodiment, from about 0.01 to about 0.03 μg per gram of cell; A specific ethanol productivity of L / day / gram (cells) would require a Ni feed rate of about 0.02 μg per minute per gram of cells;
W : In one aspect, from about 0.035 to about 0.80 μg per gram of cells per gram, in another aspect, from about 0.035 to about 0.65 μg per gram of cells, in another aspect, from about 0.035 per gram of cells per minute. To about 0.47 μg, in another embodiment, from about 0.035 to about 0.30 μg per gram of cells per gram, in another embodiment, from about 0.075 to about 0.55 μg per gram of cells, in another embodiment, per gram of cells. From about 0.075 to about 0.40 μg per minute, in another embodiment, from about 0.155 to about 0.55 μg per gram of cell, in another embodiment, from about 0.155 to about 0.40 μg per gram of cell; A specific ethanol productivity of L / day / gram (cells) would require a W feed rate of about 0.29 μg per minute per gram of cells;
Se : In one aspect, from about 0.001 to about 0.03 μg per gram of cells per gram, in another aspect, from about 0.035 to about 0.65 μg per gram of cells, in another aspect, from about 0.035 per gram of cells per minute. To about 0.47 μg, in another embodiment, from about 0.035 to about 0.30 μg per gram of cells per gram, in another embodiment, from about 0.075 to about 0.55 μg per gram of cells, in another embodiment, per gram of cells. From about 0.075 to about 0.40 μg per minute, in another embodiment, from about 0.155 to about 0.55 μg per gram of cell, in another embodiment, from about 0.155 to about 0.40 μg per gram of cell; A specific ethanol productivity of L / day / gram (cells) would require a Se feed rate of about 0.01 μg per minute per gram of cells.
別の態様では、発酵培地は、細胞1グラム当たり毎分約0.006μg以上のN、細胞1グラム当たり毎分約0.025μg以上のP、及び細胞1グラム当たり毎分約0.001μg以上のKを与えられる。この態様では、発酵培地は、下記量の下記元素の1種以上を含み得る:
N:一態様では、細胞1グラム当たり毎分約0.006〜約0.12μg、別の態様では、細胞1グラム当たり毎分約0.006〜約0.095μg、別の態様では、細胞1グラム当たり毎分約0.006〜約0.07μg、別の態様では、細胞1グラム当たり毎分約0.006〜約0.045μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.085μg、別の態様では、細胞1グラム当たり毎分約0.01〜約0.06μg、別の態様では、細胞1グラム当たり毎分約0.02〜約0.085μg、別の態様では、細胞1グラム当たり毎分約0.02〜約0.06μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.044μgのN供給速度を必要とするであろう;
P:一態様では、細胞1グラム当たり毎分約0.025〜約0.55μg、別の態様では、細胞1グラム当たり毎分約0.025〜約0.45μg、別の態様では、細胞1グラム当たり毎分約0.025〜約0.35μg、別の態様では、細胞1グラム当たり毎分約0.025〜約0.20μg、別の態様では、細胞1グラム当たり毎分約0.05〜約0.38μg、別の態様では、細胞1グラム当たり毎分約0.05〜約0.27μg、別の態様では、細胞1グラム当たり毎分約0.1〜約0.38μg、別の態様では、細胞1グラム当たり毎分約0.1〜約0.3μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.2μgのP供給速度を必要とするであろう;
K:一態様では、細胞1グラム当たり毎分約0.001〜約25μg、別の態様では、細胞1グラム当たり毎分約0.001〜約0.03μg、別の態様では、細胞1グラム当たり毎分約0.001〜約0.025μg、別の態様では、細胞1グラム当たり毎分約0.001〜約0.02μg、別の態様では、細胞1グラム当たり毎分約0.001〜約0.01μg 、別の態様では、細胞1グラム当たり毎分約0.003〜約0.02μg、別の態様では、細胞1グラム当たり毎分約0.003〜約0.015μg、別の態様では、約0.005〜約0.02μg、別の態様では、細胞1グラム当たり毎分約0.005〜約0.015μg;一例として、3g/L/日/グラム(細胞)の特定エタノール生産性は、細胞1グラム当たり毎分約0.01μgのK供給速度を必要とするであろう。
In another aspect, the fermentation medium provides about 0.006 μg or more N per gram of cells, about 0.025 μg or more of P per gram of cells, and K of about 0.001 μg or more per gram of cells. It is done. In this aspect, the fermentation medium can include one or more of the following elements in the following amounts:
N : In one aspect, from about 0.006 to about 0.12 μg per gram of cells per gram, in another aspect, from about 0.006 to about 0.095 μg per gram of cells, in another aspect, from about 0.006 per gram of cells per minute. To about 0.07 μg, in another embodiment, from about 0.006 to about 0.045 μg per gram of cells per gram, in another embodiment, from about 0.01 to about 0.085 μg per gram of cells, in another embodiment, per gram of cells. From about 0.01 to about 0.06 μg per minute, in another embodiment, from about 0.02 to about 0.085 μg per gram of cells, in another embodiment, from about 0.02 to about 0.06 μg per gram of cells; A specific ethanol productivity of L / day / gram (cells) would require an N feed rate of about 0.044 μg per minute per gram of cells;
P : In one aspect, from about 0.025 to about 0.55 μg per gram of cells per gram, in another aspect, from about 0.025 to about 0.45 μg per gram of cells, in another aspect, from about 0.025 per gram of cells per minute. To about 0.35 μg, in another embodiment, from about 0.025 to about 0.20 μg per gram of cells per gram, in another embodiment, from about 0.05 to about 0.38 μg per gram of cells, in another embodiment, per gram of cells From about 0.05 to about 0.27 μg per minute, in another embodiment from about 0.1 to about 0.38 μg per gram of cells, in another embodiment, from about 0.1 to about 0.3 μg per gram of cells per minute; A specific ethanol productivity of L / day / gram (cells) will require a P feed rate of about 0.2 μg per minute per gram of cells;
K : In one aspect, from about 0.001 to about 25 μg per gram of cells per gram, in another aspect, from about 0.001 to about 0.03 μg per gram of cells, in another aspect, from about 0.001 to about 0.001 per minute per gram of cells. About 0.025 μg, in another embodiment about 0.001 to about 0.02 μg per gram of cells per gram, in another embodiment about 0.001 to about 0.01 μg per gram of cells per minute, in another embodiment, per gram of cells From about 0.003 to about 0.02 μg per minute, in another embodiment, from about 0.003 to about 0.015 μg per gram of cells, in another embodiment, from about 0.005 to about 0.02 μg per minute, in other embodiments, from about 0.005 to about 0.02 μg per gram of cells As an example, a specific ethanol productivity of 3 g / L / day / gram (cells) would require a K feed rate of about 0.01 μg per minute per gram of cells.
別の態様では、発酵培地は約0.02質量%未満のNaHCO3、別の態様では、約0.01質量%未満のNaHCO3、別の態様では、約0.005質量%未満のNaHCO3を含む。pH調整のためNaHCO3の代わりにNH4OHを利用してよい。低ホスファートレベルのみ又はNaHCO3の利用低減との組み合わせが、より低い培地伝導率をもたらす。培地伝導率低下には、上述したよう低い希釈度及び水要求低減が必要である。関連態様では、発酵培地は約4.2〜約4.8のpHを有する。
CO供給速度は、標準立方フィート毎分(standard cubic feet per minute)(SCFM)又は標準立法フィート毎時毎リットル(standard cubic feet per hour per liter)で表すことができる。この態様では、標準立法フィート毎時毎リットルは、約0.9〜約2.0SCFM(0.025〜0.057m3/分)、別の態様では、約1.25〜約1.75SCFM(0.0354〜0.0496m3/分)の範囲にあり得る。別の態様では、平均CO供給速度は、約0.016:1〜約0.04:1、別の態様では、約0.02:1〜約0.04:1、別の態様では、約0.02:1〜約0.035:1、別の態様では、約0.025:1〜約0.035:1、別の態様では、約0.025:1〜約0.03:1のCO供給速度対発酵槽容積の比を維持するのに有効なCO供給速度である。
In another aspect, the fermentation medium comprises less than about 0.02% by weight NaHCO 3 , in another aspect, less than about 0.01% by weight NaHCO 3 , in another aspect, less than about 0.005% by weight NaHCO 3 . NH 4 OH may be used instead of NaHCO 3 for pH adjustment. Only a low phosphate level or a combination with reduced utilization of NaHCO 3 results in lower media conductivity. To reduce the medium conductivity, it is necessary to reduce the dilution and water requirement as described above. In a related embodiment, the fermentation medium has a pH of about 4.2 to about 4.8.
The CO feed rate can be expressed in standard cubic feet per minute (SCFM) or standard cubic feet per hour per liter. In this embodiment, standard cubic feet per hour liters range from about 0.9 to about 2.0 SCFM (0.025 to 0.057 m 3 / min), and in another embodiment about 1.25 to about 1.75 SCFM (0.0354 to 0.0496 m 3 / min). Can be. In another embodiment, the average CO feed rate is from about 0.016: 1 to about 0.04: 1, in another embodiment, from about 0.02: 1 to about 0.04: 1, in another embodiment, from about 0.02: 1 to about 0.035: 1. In another embodiment, the CO feed rate effective to maintain a ratio of CO feed rate to fermentor volume of about 0.025: 1 to about 0.035: 1, and in another embodiment about 0.025: 1 to about 0.03: 1. It is.
別の態様では、プロセスは、H2転化率をモニターする工程及び約25%以上、別の態様では、約25%〜約95%、別の態様では、約30%〜約90%、別の態様では、約35%〜約85%、別の態様では、約40%〜約80%、別の態様では、約40%〜約70%、別の態様では、約40%〜約60%、別の態様では、約40%〜約50%のH2転化率を維持する工程を含む。プロセスは、CO取込みをモニターする工程及び約0.001〜約10ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.001〜約5ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.001〜約4ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.001〜約3ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.001〜約2ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.001〜約1ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約9ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約5ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約4ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約3ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約2ミリモル/分/グラム(乾燥細胞)、別の態様では、約0.05〜約1ミリモル/分/グラム(乾燥細胞)、別の態様では、約1〜約8ミリモル/分/グラム(乾燥細胞)、別の態様では、約1〜約5ミリモル/分/グラム(乾燥細胞)、別の態様では、約1〜約4ミリモル/分/グラム(乾燥細胞)、別の態様では、約1〜約3ミリモル/分/グラム(乾燥細胞)、別の態様では、約1〜約2ミリモル/分/グラム(乾燥細胞)のCO取込みを維持する工程をさらに含んでよい。 In another embodiment, the process, H 2 conversion monitor to process and about 25% or more, in another embodiment, from about 25% to about 95%, in another embodiment, from about 30% to about 90%, of another In embodiments, from about 35% to about 85%, in other embodiments from about 40% to about 80%, in other embodiments, from about 40% to about 70%, in other embodiments, from about 40% to about 60%, in another aspect, comprising the step of maintaining of H 2 conversion of about 40% to about 50%. The process includes monitoring CO uptake and from about 0.001 to about 10 mmol / min / gram (dry cells), in another embodiment, from about 0.001 to about 5 mmol / min / gram (dry cells), in another embodiment, About 0.001 to about 4 mmol / min / gram (dry cells), in another embodiment about 0.001 to about 3 mmol / min / gram (dry cells), in another embodiment about 0.001 to about 2 mmol / min / gram (Dry cells), in another embodiment, from about 0.001 to about 1 mmol / min / gram (dry cells), in another embodiment, from about 0.05 to about 9 mmol / min / gram (dry cells), in another embodiment, About 0.05 to about 5 mmol / min / gram (dry cells), in another embodiment about 0.05 to about 4 mmol / min / gram (dry cells), in another embodiment about 0.05 to about 3 mmol / min / gram (Dry cells), in another embodiment, from about 0.05 to about 2 mmol / min / gram (dry cells), in another embodiment, from about 0.05 to about 1 mmol / min / gram (dry cells), in another embodiment About 1 to about 8 mmol / min / gram (dry cells), in another embodiment about 1 to about 5 mmol / min / gram (dry cells), in another embodiment about 1 to about 4 mmol / min / gram / dry Maintain CO uptake of grams (dry cells), in another embodiment, about 1 to about 3 mmol / min / gram (dry cells), in another embodiment, about 1 to about 2 mmol / min / gram (dry cells) The process of carrying out may be further included.
実施例1:適合性試験
以前に利用した微量金属溶液は下記成分を含んだ(全てグラム/リットルで表した)。
原液 適合性試験
1) ZnSO4 *7H2O 0.5222 2.35
2) COCl2 *6H2O 1.6 7.196
3) NiCl2 *6H2O 0.4944 2.222
4) Na2SeO3 0.16 0.72
5) Na2WO4 *2H2O 3.2 14.404
6) H3PO4(85%) 10% N/A
Example 1 : Compatibility test The previously used trace metal solution contained the following components (all expressed in grams / liter):
Stock solution compatibility test
1) ZnSO 4 * 7H 2 O 0.5222 2.35
2) COCl 2 * 6H 2 O 1.6 7.196
3) NiCl 2 * 6H 2 O 0.4944 2.222
4) Na 2 SeO 3 0.16 0.72
5) Na 2 WO 4 * 2H 2 O 3.2 14.404
6) H 3 PO 4 (85%) 10% N / A
1つの溶液に5種より多くの微量金属全てを維持するためには酸性マトリックスが必要である。しかしながら、これらの金属はそれらだけで水に高溶解性である。そこで、後述するように適合性試験を行なった。各微量金属の個々の溶液を作製した。各溶液の濃度は、原液中の各微量金属の濃度に等しかった。各溶液を互いに混合し、室温で一晩インキュベートした。翌朝の溶液を濁度について目視検査し、分光光度計で(ボルテックス)溶液の光学密度を測定した。結果を以下に示す。 An acidic matrix is required to maintain all of the more than five trace metals in one solution. However, these metals are highly soluble in water by themselves. Therefore, a compatibility test was performed as described later. Individual solutions of each trace metal were made. The concentration of each solution was equal to the concentration of each trace metal in the stock solution. Each solution was mixed with each other and incubated overnight at room temperature. The next morning solution was visually inspected for turbidity and the optical density of the (vortex) solution was measured with a spectrophotometer. The results are shown below.
上記データは、Zn、Co及びNiと沈殿を形成しないようにSe及びWをプロトン化した状態にしておくためには酸性マトリックスが必要であることを示している。上記知見に基づいて、1つの微量金属原液の代わりに2つの微量金属原液を作製した。第1原液にはZn、Co、及びNiを含め、第2原液にはW及びSeを含めた。この調製方法はH3PO4の使用を低減する。実験原液中のリン酸の完全な排除を補うため、第1原液に添加するリン酸の量を0.075ml/Lから0.2ml/Lに増やした。従って、培地中のH3PO4の全体的な正味の低減は76%だった。 The above data shows that an acidic matrix is required to keep Se and W protonated so as not to form precipitates with Zn, Co and Ni. Based on the above findings, two trace metal stock solutions were prepared instead of one trace metal stock solution. The first stock solution contained Zn, Co, and Ni, and the second stock solution contained W and Se. This preparation method reduces the use of H 3 PO 4 . To compensate for the complete elimination of phosphoric acid in the experimental stock solution, the amount of phosphoric acid added to the first stock solution was increased from 0.075 ml / L to 0.2 ml / L. Therefore, the overall net reduction of H 3 PO 4 in the medium was 76%.
実施例2:低減ホスファート培地の使用
上記(76%少ないリン酸を含有する)培地を定常状態培養について以下のように4段階で試験した。
1. 定常培養に存在する培地を変性培地と交換した(T=0時間)。
2. 成長培地中のNH4ClをNH4OHと交換した。予防策としてH2SO4を成長培地に加えて反応器のpHを4.5で維持した(T=108.74時間)。
3. H2SO4を培地から除去し、塩基としてNaHCO3をNH4OHと交換して反応器のpHを制御した(T=158.42時間)。
4. 第1原液中の成分を発酵培地に直接加えた(T=489.07時間)。
図1は、低ホスファート培地及びpHを制御し、かつ窒素源として作用するための塩基としてのNH4OHの使用に関する定常状態クロストリジウム・リュングダリイ培養の性能を示す。発酵中の事象は以下のとおりだった。
Example 2 Use of Reduced Phosphate Medium The above medium (containing 76% less phosphate) was tested for steady state culture in four stages as follows.
1. The medium present in stationary culture was replaced with denatured medium (T = 0 hour).
2. NH 4 Cl in the growth medium was exchanged with NH 4 OH. As a precaution, H 2 SO 4 was added to the growth medium to maintain the reactor pH at 4.5 (T = 108.74 hours).
3. H 2 SO 4 was removed from the medium and the pH of the reactor was controlled by exchanging NaHCO 3 as base with NH 4 OH (T = 158.42 hours).
4. The ingredients in the first stock solution were added directly to the fermentation medium (T = 489.07 hours).
FIG. 1 shows the performance of a steady-state Clostridium lünddalii culture for low phosphate medium and the use of NH 4 OH as a base to control pH and act as a nitrogen source. Events during fermentation were as follows.
108.74時間には、0.35ml/LのH2SO4(75%)を含有する培地を反応器に添加した。NH4Clを培地から除去し、NH4OHポンプ処理を開始した。これは、反応器にポンプ処理される追加塩基がpH設定点を飛び越えないようにするために行なった。このpHではH3PO4は一塩基性であり、H2SO4は二塩基性であることを考慮して、H3PO4として排除されるプロトンの量に基づいて添加量を計算した。後に培養は、依然として塩基を使用し、H2SO4は排除されていることを確証した。
135.32時間に開始して、NaHCO3の塩基溶液をNH4OHに交換した。0.5Mの最終溶液に決めるまで、NH4OHポンプ処理の流速にとともに塩基の濃度を調整した。この濃度を用いると、培養に窒素を与えるために追加のNH4OHを加える必要がなかった。
279.32時間と441.82時間の間に、培地中のビタミン濃度を1.6ml/Lに上昇させた。
392.22時間には、細胞密度を3g/Lに低減した。
最終培地組成変更を489.07時間に加えた。これは、第1原液成分をそれらの固体形態で(全ての他の成分で行なうように)直接培地に添加することによって行なった。第2原液成分は水溶液として添加した。
本発明を特定の実施形態、その実施例及び応用を利用して開示したが、当業者は、特許請求の範囲に明記する本発明の範囲を逸脱することなく、それらに多くの変更形態及び変形形態を加えることができるであろう。
At 108.74 hours, medium containing 0.35 ml / L H 2 SO 4 (75%) was added to the reactor. NH 4 Cl was removed from the medium and NH 4 OH pumping was started. This was done to prevent additional base pumped into the reactor from jumping over the pH set point. Considering that H 3 PO 4 is monobasic and H 2 SO 4 is dibasic at this pH, the amount added was calculated based on the amount of protons excluded as H 3 PO 4 . Later cultures still used base and confirmed that H 2 SO 4 was eliminated.
Starting at 135.32 hours, the base solution of NaHCO 3 was replaced with NH 4 OH. The base concentration was adjusted with the NH 4 OH pumping flow rate until a final 0.5 M solution was determined. With this concentration, it was not necessary to add additional NH 4 OH to feed the culture with nitrogen.
Between 279.32 hours and 441.82 hours, the vitamin concentration in the medium was increased to 1.6 ml / L.
At 392.22 hours, the cell density was reduced to 3 g / L.
A final media composition change was added at 489.07 hours. This was done by adding the first stock solution components in their solid form directly (as is done with all other components) to the medium. The second stock solution component was added as an aqueous solution.
While the invention has been disclosed using specific embodiments, examples thereof and applications, those skilled in the art will recognize many modifications and variations thereto without departing from the scope of the invention as set forth in the claims. A form could be added.
Claims (27)
少なくとも1種の遷移金属元素を含む液体培地を、少なくとも1種の他の遷移金属元素及び任意選択的に1種の非金属元素を含む液体培地とブレンドして発酵培地を与える工程;
CO含有基質を前記発酵培地と接触させる工程;及び
前記CO含有基質を発酵させて酸性pHを与える工程
を含む発酵プロセスであって、
前記プロセスが、1種以上の遷移金属元素と1種以上の非金属元素との沈殿を防止するのに有効であり、かつ
生成されるエタノール1米国ガロン(3.8リットル)当たり前記発酵培地に与えられる約2米国ガロン(7.6リットル)以下の水を利用するのに有効である、プロセス。 The following process:
Blending a liquid medium comprising at least one transition metal element with a liquid medium comprising at least one other transition metal element and optionally one non-metallic element to provide a fermentation medium;
Contacting the CO-containing substrate with the fermentation medium; and fermenting the CO-containing substrate to provide an acidic pH comprising:
The process is effective to prevent precipitation of one or more transition metal elements and one or more non-metallic elements and is fed to the fermentation medium per 1 US gallon (3.8 liters) of ethanol produced A process that is effective to utilize less than about 2 US gallons (7.6 liters) of water.
前記CO含有基質を発酵槽に与える工程及び前記CO含有基質を発酵培地と接触させる工程(前記発酵培地は、Zn、Co及びNiから成る群より選択される1種以上の元素を含む第1溶液を、W及びSeから成る群より選択される1種以上の元素を含む第2溶液と、30mS/cm以下の伝導率を有し、3mM以下のホスファートを含む発酵培地を与えるのに有効な量でブレンドする工程を含むプロセスによって与えられる);及び
前記CO含有基質を発酵させる工程
を含む発酵プロセスであって、
前記プロセスが、10gの全アルコール/(L・日)以上のSTYを与えるのに有効であり、
前記発酵プロセスが、生成されるエタノール1米国ガロン(3.8リットル)当たり前記発酵培地に与えられる約2米国ガロン(7.6リットル)以下の水を利用するのに有効である、プロセス。 A fermentation process for a CO-containing substrate comprising the following steps:
A step of supplying the CO-containing substrate to a fermentor and a step of bringing the CO-containing substrate into contact with a fermentation medium (the fermentation medium is a first solution containing one or more elements selected from the group consisting of Zn, Co and Ni) A second solution containing one or more elements selected from the group consisting of W and Se, and an amount effective to provide a fermentation medium having a conductivity of 30 mS / cm or less and a phosphate of 3 mM or less. And a fermentation process comprising the step of fermenting said CO-containing substrate,
The process is effective to give 10 g total alcohol / (L · day) or more STY,
A process wherein the fermentation process is effective to utilize no more than about 2 US gallons (7.6 liters) of water fed to the fermentation medium per 1 US gallon (3.8 liters) of ethanol produced.
細胞1グラム当たり毎分約0.04μg以上のZn、
細胞1グラム当たり毎分約0.018μg以上のCo、
細胞1グラム当たり毎分約0.02μg以上のNi、
細胞1グラム当たり毎分約0.29μg以上のW、及び
細胞1グラム当たり毎分約0.01μg以上のSe
の少なくとも1つ以上が与えられる、請求項11の発酵プロセス。 In the fermentation medium, about 0.04 μg or more of Zn per minute per gram of the following cells,
About 0.018 μg or more of Co per minute per gram of cells,
About 0.02 μg or more of Ni per minute per gram of cells,
More than about 0.29 μg W per gram per cell and more than about 0.01 μg Se per gram per cell
12. The fermentation process of claim 11, wherein at least one or more of is provided.
細胞1グラム当たり約0.044μg以上の窒素、
細胞1グラム当たり約0.2μg以上のリン、又は
細胞1グラム当たり約0.01μg以上のカリウム
の少なくとも1つ以上が与えられる、請求項11の発酵プロセス。 In the fermentation medium, about 0.044 μg or more of nitrogen per gram of the following cells,
12. The fermentation process of claim 11, wherein at least one or more of about 0.2 μg or more of phosphorus per gram of cells or about 0.01 μg or more of potassium per gram of cells is provided.
Zn、Co、Niの1種以上から成る群より選択される元素を含む溶液を、W、Seの1種以上から成る群より選択される元素を含む溶液と、約30mS/cm以下の伝導率を有する発酵培地を与えるのに有効な量でブレンドする工程を含み、
前記発酵培地が、約3mMより多くのホスファートを有する発酵培地より約10%〜約40%少ない水を必要とする、前記プロセス。 A process for reducing the water used for the preparation of the fermentation medium, comprising the following steps:
A solution containing an element selected from the group consisting of one or more of Zn, Co, and Ni, a solution containing an element selected from the group consisting of one or more of W and Se, and a conductivity of about 30 mS / cm or less Blending in an amount effective to provide a fermentation medium having
The process, wherein the fermentation medium requires about 10% to about 40% less water than a fermentation medium with more than about 3 mM phosphate.
細胞1グラム当たり毎分約0.04μg以上のZn、
細胞1グラム当たり毎分約0.018μg以上のCo、
細胞1グラム当たり毎分約0.02μg以上のNi、
細胞1グラム当たり毎分約0.29μg以上のW、及び
細胞1グラム当たり毎分約0.01μg以上のSe
の少なくとも1つ以上が与えられる、請求項22の発酵プロセス。 In the fermentation medium, about 0.04 μg or more of Zn per minute per gram of the following cells,
About 0.018 μg or more of Co per minute per gram of cells,
About 0.02 μg or more of Ni per minute per gram of cells,
More than about 0.29 μg W per gram per cell and more than about 0.01 μg Se per gram per cell
23. The fermentation process of claim 22, wherein at least one or more of is provided.
細胞1グラム当たり約0.044μg以上の窒素、
細胞1グラム当たり約0.2μg以上のリン、又は
細胞1グラム当たり約0.01μg以上のカリウム
の少なくとも1つ以上が与えられる、請求項22の発酵プロセス。 In the fermentation medium, about 0.044 μg or more of nitrogen per gram of the following cells,
23. The fermentation process of claim 22, wherein at least one or more of about 0.2 μg or more of phosphorus per gram of cells or about 0.01 μg or more of potassium per gram of cells is provided.
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CN110317837B (en) | 2024-04-02 |
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CA2913196C (en) | 2022-07-19 |
CN105392890A (en) | 2016-03-09 |
TW201534718A (en) | 2015-09-16 |
WO2014200810A1 (en) | 2014-12-18 |
AU2014278580B2 (en) | 2017-08-31 |
RU2015151914A (en) | 2017-07-14 |
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